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Abstract:

The present invention relates to methods of treating, preventing, or
lessening the severity of resistant bacterial infections in mammals,
utilizing compounds of formula I or formula VII or pharmaceutically salts
thereof. The present invention also relates to methods of using compounds
of formula I or formula VII in combination with one or more additional
antibacterial agents and/or one or more additional therapeutic agents
that increase the susceptibility of bacterial organisms to antibiotics.

Claims:

1. A method of controlling, treating or reducing the advancement,
severity or effects of a nosocomial or a non-nosocomial of a bacterial
infection in a patient, wherein the bacterial infection is characterized
by the presence of one or more of the following: Methicillin resistant
Staphylococci, Fluoroquinolone resistant Staphylococci, Glycopepetide
resistant Staphylococci, Macrolide-Lincosamide-Streptogramin resistant
Staphylococci, Linezolid resistant Enterococci, Glycopepetide resistant
Enterococci, β-lactam resistant Enterococci, Penicillin resistant
Streptococci, Macrolide resistant Streptococci, Ketolide resistant
Streptococci, Fluoroquinolone resistant Streptococci, β-lactam
resistant Haemophilus, Fluoroquinolone resistant Haemophilus, Macrolide
resistant Haemophilus, Macrolide resistant Mycoplasma, Isoniazid
resistant Mycobacterium, Rifampin resistant Mycobacterium, or
β-lactam resistant Moraxella, comprising the step of administering
to said patient a compound of formula I: ##STR00580## or a
pharmaceutically acceptable salt thereof, wherein: W is selected from
nitrogen, CH, or CF; X is selected from CH or CF; Z is O or NH; R1
is phenyl or a 5-6 membered heteroaryl ring having 1-3 heteroatoms
independently selected from oxygen, nitrogen, or sulfur, wherein: R1
is substituted with 0-3 groups independently selected from -(T)y-Ar,
R', oxo, C(O)R', CO2R', OR', N(R')2, SR', NO2, halogen,
CN, C(O)N(R')2, NR'C(O)R', SO2R', SO2N(R')2, or
NR'SO2R'; y is 0 or 1; T is a straight or branched C1-4
alkylidene chain, wherein one methylene unit of T is optionally replaced
by --O--, --NH--, or --S--; each R' is independently selected from
hydrogen, C1-4 aliphatic, or a 5-6 membered saturated, unsaturated,
or aryl ring having 0-3 heteroatoms independently selected from nitrogen,
oxygen, or sulfur, wherein: R' is substituted with 0-3 groups
independently selected from halogen, oxo, Ro, N(RO)2,
ORo, CO2Ro, NRoC(O)Ro, C(O)N(RO)2,
SO2Ro, SO2N(RO)2, or NRoSO2Ro,
wherein: each Ro is independently selected from hydrogen, C1-4
aliphatic, or a 5-6 membered saturated, unsaturated, or aryl ring having
0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur,
and wherein: two substituents on adjacent positions of R1 may be
taken together to form a 5-7 membered saturated, partially unsaturated,
or aryl ring having 0-3 heteroatoms independently selected from nitrogen,
oxygen, or sulfur; Ar is a 3-8 membered saturated, unsaturated, or aryl
ring, a 3-7 membered heterocyclic ring having 1-3 heteroatoms
independently selected from nitrogen, oxygen, or sulfur, or a 5-6
membered heteroaryl ring having 1-3 heteroatoms independently selected
from nitrogen, oxygen, or sulfur, wherein: Ar is substituted with 0-3
groups independently selected from R', oxo, CO2R', OR', N(R')2,
SR', NO2, halogen, CN, C(O)N(R')2, NR'C(O)R', SO2R',
C(O)R', SO2N(R')2, or NR'SO2R'; R2 is selected from
hydrogen or a C1-3 aliphatic group; and Ring A is a 5-6 membered
heteroaryl ring having 1-4 heteroatoms independently selected from
nitrogen, oxygen, or sulfur, provided that said ring has a hydrogen-bond
acceptor in the position adjacent to the point of attachment to Ring B,
wherein: Ring A is substituted with 0-3 groups independently selected
from R', oxo, CO2R', OR', N(R')2, SR', NO2, halogen, CN,
C(O)N(R')2, NR'C(O)R', SO2R', SO2N(R')2, or
NR'SO2R', and wherein: two substituents on adjacent positions of
Ring A may be taken together to form a 5-7 membered saturated, partially
unsaturated, or aryl ring having 0-3 heteroatoms independently selected
from nitrogen, oxygen, or sulfur.

2. A method of controlling, treating or reducing the advancement,
severity or effects of a nosocomial or a non-nosocomial bacterial
infection in a patient, wherein the bacterial infection is characterized
by the presence of one or more of the following: Methicillin resistant
Staphylococci, Fluoroquinolone resistant Staphylococci, Glycopepetide
resistant Staphylococci, Macrolide-Lincosamide-Streptogramin resistant
Staphylococci, Linezolid resistant Enterococci, Glycopepetide resistant
Enterococci, β-lactam resistant Enterococci, Penicillin resistant
Streptococci, Macrolide resistant Streptococci, Ketolide resistant
Streptococci, Fluoroquinolone resistant Streptococci, β-lactam
resistant Haemophilus, Fluoroquinolone resistant Haemophilus, Macrolide
resistant Haemophilus, Macrolide resistant Mycoplasma, Isoniazid
resistant Mycobacterium, Rifampin resistant Mycobacterium, or
β-lactam resistant Moraxella, comprising the step of administering
to said patient a compound of formula VII: ##STR00581## or a
pharmaceutically acceptable salt thereof, wherein: V is selected from
nitrogen, CH, or CF; R3 is hydrogen or C1-4 aliphatic, wherein:
when R3 is C1-4 aliphatic, R3 is substituted with 0-3
groups independently selected from OH, R5, or OR5; wherein:
R5 is C1-3 aliphatic, wherein: two R5 aliphatic groups may
be optionally taken together with the carbon to which they are bound to
form a C3-4 cycloalkyl ring; provided that if R3 is hydrogen,
then V is not nitrogen or CH; R4 is a C1-3 aliphatic group; and
Ring C is a 6-membered heteroaryl ring having 1-2 nitrogens, wherein:
Ring C is substituted with 1-3 groups selected from R6; wherein:
each R6 is independently selected from OR7 or halogen; and
R7 is C1-4 aliphatic; or Ring C is an unsubstituted
2-pyrimidine ring.

3. The method according to claim 2, comprising the step of administering
to said patient a compound of formula VIIA, VIIB, or VIIC: ##STR00582##
or a pharmaceutically acceptable salt thereof, wherein: R4 is a
C1-3 aliphatic group; and Ring C is a 6-membered heteroaryl ring
having 1-2 nitrogens, wherein: Ring C is substituted with 1-3 groups
selected from R6; wherein: each R6 is independently selected
from OR7 or halogen; R7 is C1-4 aliphatic; or Ring C is an
unsubstituted 2-pyrimidine ring.

25. The method according to claim 1, further comprising the step of
administering to said human one or more additional therapeutic agents
either as part of a multiple dosage form together with said compound or
as a separate dosage form wherein said one or more additional therapeutic
agents include an antibiotic selected from a natural penicillin, a
penicillinase-resistant penicillin, an antipseudomonal penicillin, an
aminopenicillin, a first generation cephlosporin, a second generation
cephalosporin, a third generation cephalosporin, a fourth generation
cephalosporin, a carbapenem, a cephamycin, a monobactam, a quinolone, a
fluoroquinolone, an aminoglycoside, a macrolide, a ketolide, a
tetracycline, a glycopeptide, a streptogramin, an oxazolidone, a
rifamycin, or other antibiotics.

26. The method according to claim 25, wherein said natural penicillin is
Benzathine penicillin G, Penicillin G or Penicillin V, said
penicillinase-resistant penicillin is Cloxacillin, Dicloxacillin,
Nafcillin or Oxacillin, said antipseudomonal penicillin is Carbenicillin,
Mezlocillin, Pipercillin, Pipercillin/tazobactam, Ticaricillin or
Ticaricillin/Clavulanate, said aminopenicillin is Amoxicillin, Ampicillin
or Ampicillin/Sulbactam, said first generation cephalosporin is
Cefazolin, Cefadroxil, Cephalexin or Cephadrine, said second generation
cephalosporin is Cefaclor, Cefaclor-CD, Cefamandole, Cefonacid,
Cefprozil, Loracarbef or Cefuroxime, said third generation cephalosporin
is Cefdinir, Cefixime, Cefoperazone, Cefotaxime, Cefpodoxime,
Ceftazidime, Ceftibuten, Ceftizoxme or Ceftriaxone, said fourth
generation cephalosporin is Cefepime, said Cephamycin is Cefotetan or
Cefoxitin, said carbapenem is Imipenem or Meropenem, said monobactam is
Aztreonam, said quinolone is Cinoxacin, Nalidixic acid, Oxolininc acid or
Pipemidic acid, said fluoroquinolone is Cirpofloxacin, Enoxacin,
Gatifloxacin, Grepafloxacin, Levofloxacin, Lomefloxacin, Moxifloxacin,
Norfloxacin, Ofloxacin or Sparfloxacin, said aminoglycoside is Amikacin,
Gentamicin, Kanamycin, Neomycin, Netilmicin, Spectinomycin, Streptomycin
or Tobramycin, said macrolide is Azithromycin, Clarithromycin or
Erythromycin, said ketolide is Telithromycin, said Tetracycline is
Chlortetracycline, Demeclocycline, Doxycycline, Minocycline or
Tetracycline, said glycopeptide is Oritavancin, Teicoplanin or
Vancomycin, said streptogramin is Dalfopristin/quinupristin, said
oxazolidone is Linezolid, said Rifamycin is Rifabutin or Rifampin and
said other antibiotic is bactitracin, chloramphenicol, clindamycin,
isoniazid, metronidazole, polymyxin B, pyrazinamide, or
trimethoprim/sulfamethoxazole.

27. The method according to claim 26 wherein said natural penicillin is
Penicillin G, said penicillinase-resistant penicillin is Nafcillin or
Oxacillin, said antipseudomonal penicillin is Pipercillin/tazobactam,
said aminopenicillin is Amoxicillin, said first generation cephalosporin
is Cephalexin, said second generation cephalosporin is Cefaclor,
Cefaclor-CD or Cefuroxime, said third generation cephalosporin is
Ceftazidime or Ceftriaxone, said fourth generation cephalosporin is
Cefepime, said fluoroquinolone is Cirpofloxacin, Gatifloxacin,
Levofloxacin or Moxifloxacin, said aminoglycoside is Tobramycin, said
macrolide is Azithromycin or Clarithromycin, said Tetracycline is
Doxycycline, said glycopeptide is Vancomycin, said Rifamycin is Rifampin
and said other antibiotic is isoniazid, pyrazinamide, or
trimethoprim/sulfamethoxazole.

[0002] This invention is in the field of medicinal chemistry and relates
to compounds, and pharmaceutical compositions thereof, that inhibit
bacterial gyrase and Topo IV. The compounds are useful as inhibitors of
bacterial gyrase and Topo IV activity. The present invention also relates
to methods of using the compounds of this invention for treating
resistant bacterial infections in patients. The present invention also
relates to treating resistant bacterial infections in patients using
compounds of the present invention in combination with one or more
additional antibacterial agents and/or one or more additional therapeutic
agents that increase the susceptibility of bacterial organisms to
antibiotics.

BACKGROUND OF THE INVENTION

[0003] Bacterial resistance to antibiotics has long been recognized, and
it is today considered to be a serious worldwide health problem. As a
result of resistance, some bacterial infections are either difficult to
treat with antibiotics or even untreatable. This problem has become
especially serious with the recent development of multiple drug
resistance in certain strains of bacteria, such as Streptococcus
pneumoniae (SP), Mycobacterium tuberculosis, and Enterococcus. The
appearance of vancomycin resistant enterococcus was particularly alarming
because vancomycin was formerly the only effective antibiotic for
treating this infection, and had been considered for many infections to
be the drug of "last resort". While many other drug-resistant bacteria do
not cause life-threatening disease, such as enterococci, there is the
fear that the genes which induce resistance might spread to more deadly
organisms such as Staphylococcus aureus, where methicillin resistance is
already prevalent (De Clerq, et al., Current Opinion in Anti-infective
Investigational Drugs, 1999, 1, 1; Levy, "The Challenge of Antibiotic
Resistance", Scientific American, March, 1998).

[0004] Another concern is how quickly antibiotic resistance can spread.
For example, until the 1960's SP was universally sensitive to penicillin,
and in 1987 only 0.02% of the SP strains in the U.S. were resistant.
However, by 1995 it was reported that SP resistance to penicillin was
about seven percent and as high as 30% in some parts of the U.S. (Lewis,
FDA Consumer magazine (September, 1995); Gershman in The Medical
Reporter, 1997).

[0005] Hospitals, in particular, serve as centers for the formation and
transmission of drug-resistant organisms. Infections occurring in
hospitals, known as nosocomial infections, are becoming an increasingly
serious problem. Of the two million Americans infected in hospitals each
year, more than half of these infections resist at least one antibiotic.
The Center for Disease Control reported that in 1992, over 13,000
hospital patients died of bacterial infections that were resistant to
antibiotic treatment (Lewis, "The Rise of Antibiotic-Resistant
Infections", FDA Consumer magazine, September, 1995).

[0006] As a result of the need to combat drug-resistant bacteria and the
increasing failure of the available drugs, there has been a resurgent
interest in discovering new antibiotics. One attractive strategy for
developing new antibiotics is to inhibit DNA gyrase, a bacterial enzyme
necessary for DNA replication, and therefore, necessary for bacterial
cell growth and division. Gyrase activity is also associated with events
in DNA transcription, repair and recombination.

[0007] Gyrase is one of the topoisomerases, a group of enzymes which
catalyze the interconversion of topological isomers of DNA (see
generally, Kornberg and Baker, DNA Replication, 2d Ed., Chapter 12, 1992,
W.H. Freeman and Co.; Drlica, Molecular Microbiology, 1992, 6, 425;
Drlica and Zhao, Microbiology and Molecular Biology Reviews, 1997, 61,
377). Gyrase itself controls DNA supercoiling and relieves topological
stress that occurs when the DNA strands of a parental duplex are
untwisted during the replication process. Gyrase also catalyzes the
conversion of relaxed, closed circular duplex DNA to a negatively
superhelical form which is more favorable for recombination. The
mechanism of the supercoiling reaction involves the wrapping of gyrase
around a region of the DNA, double strand breaking in that region,
passing a second region of the DNA through the break, and rejoining the
broken strands. Such a cleavage mechanism is characteristic of a type II
topoisomerase. The supercoiling reaction is driven by the binding of ATP
to gyrase. The ATP is then hydrolyzed during the reaction. This ATP
binding and subsequent hydrolysis cause conformational changes in the
DNA-bound gyrase that are necessary for its activity. It has also been
found that the level of DNA supercoiling (or relaxation) is dependent on
the ATP/ADP ratio. In the absence of ATP, gyrase is only capable of
relaxing supercoiled DNA.

[0008] Bacterial DNA gyrase is a 400 kilodalton protein tetramer
consisting of two A (GyrA) and two B subunits (GyrB). Binding and
cleavage of the DNA is associated with GyrA, whereas ATP is bound and
hydrolyzed by the GyrB protein. GyrB consists of an amino-terminal domain
which has the ATPase activity, and a carboxy-terminal domain which
interacts with GyrA and DNA. By contrast, eukaryotic type II
topoisomerases are homodimers that can relax negative and positive
supercoils, but cannot introduce negative supercoils. Ideally, an
antibiotic based on the inhibition of bacterial DNA gyrase would be
selective for this enzyme and be relatively inactive against the
eukaryotic type II topoisomerases.

[0009] The widely used quinolone antibiotics inhibit bacterial DNA gyrase.
Examples of the quinolones include the early compounds such as nalidixic
acid and oxolinic acid, as well as the later, more potent
fluoroquinolones such as norfloxacin, ciprofloxacin, and trovafloxacin.
These compounds bind to GyrA and stabilize the cleaved complex, thus
inhibiting overall gyrase function, leading to cell death. However, drug
resistance has also been recognized as a problem for this class of
compounds (WHO Report, "Use of Quinolones in Food Animals and Potential
Impact on Human Health", 1998). With the quinolones, as with other
classes of antibiotics, bacteria exposed to earlier compounds often
quickly develop cross-resistance to more potent compounds in the same
class.

[0010] There are fewer known inhibitors that bind to GyrB. Examples
include the coumarins, novobiocin and coumermycin Al, cyclothialidine,
cinodine, and clerocidin. The coumarins have been shown to bind to GyrB
very tightly. For example, novobiocin makes a network of hydrogen bonds
with the protein and several hydrophobic contacts. While novobiocin and
ATP do appear to bind within the ATP binding site, there is minimal
overlap in the bound orientation of the two compounds. The overlapping
portions are the sugar unit of novobiocin and the ATP adenine (Maxwell,
Trends in Microbiology, 1997, 5, 102).

[0011] For coumarin-resistant bacteria, the most prevalent point mutation
is at a surface arginine residue that binds to the carbonyl of the
coumarin ring (Arg136 in E. coli GyrB). While enzymes with this mutation
show lower supercoiling and ATPase activity, they are also less sensitive
to inhibition by coumarin drugs (Maxwell, Mol. Microbiol., 1993, 9, 681).

[0012] Despite being potent inhibitors of gyrase supercoiling, the
coumarins have not been widely used as antibiotics. They are generally
not suitable due to their low permeability in bacteria, eukaryotic
toxicity, and poor water solubility (Maxwell, Trends in Microbiology,
1997, 5, 102). It would be desirable to have a new, effective GyrB
inhibitor that overcomes these drawbacks. Such an inhibitor would be an
attractive antibiotic candidate, without a history of resistance problems
that plague other classes of antibiotics.

[0013] Replication fork movement along circular DNA can generate
topological changes both ahead of the replication complex as well as
behind in the already replicated regions (Champoux, J. J., Annu. Rev.
Biochem., 2001, 70, 369-413). While DNA gyrase can introduce negative
supercoils to compensate for the topological stresses ahead of the
replication fork, some overwinding can diffuse back into the already
replicated region of DNA resulting in precatenanes. If not removed, the
presence of the precatenanes can result in interlinked (catenated)
daughter molecules at the end of replication. TopoIV is responsible for
separating the catenated daughter plasmids as well as removal of
precatenanes formed during replication ultimately allowing for
segragation of the daughter molecules into daughter cells. Topo IV is
composed of two ParC and 2 parE subunits as a C2E2 tetramer (where the C
and E monomers are homologuous to the A and B monomers of gyrase,
respectively) that requires ATP hydrolysis (at the N-terminus of the E
subunit) to reset the enzyme to re-enter the catalytic cycle. Topo IV is
highly conserved among bacteria and is essential for bacterial
replication (Drlica and Zhao, Microbiol. Mol. Biol. Rev., 1997, 61, 377).

[0014] While little attention has been paid to inhibitors that target ParE
of TopoIV, the action of the newer quinolones on the ParC region has been
widely studied (Hooper, D. C., Clin. Infect. Dis., 2000, 31(Suppl 2):
S24-28). It has been demonstrated that moxifloxacin and gatifloxacin have
more balanced activities against Gyrase and TopoIV resulting in expanded
Gram positive coverage as well as lower levels of resistance caused
primary-target mutation. In those cases, susceptibility is limited by the
sensitivity of the second target to the antibacterial agent. Thus, agents
that can effectively inhibit multiple essential targets can result in an
expanded spectrum of potencies, improved antibacterial potencies,
improved potency against single target mutants, and/or lower spontaneous
rates of resistance.

[0015] As bacterial resistance to antibiotics has become an important
public health problem, there is a continuing need to develop newer and
more potent antibiotics. More particularly, there is a need for
antibiotics that represent a new class of compounds not previously used
to treat bacterial infection. Such compounds would be particularly useful
in treating nosocomial infections in hospitals where the formation and
transmission of resistant bacteria are becoming increasingly prevalent.

SUMMARY OF THE INVENTION

[0016] It has now been found that compounds of this invention are useful
in methods of treating, preventing, or lessening the severity of a
bacterial infection in a patient, wherein the bacterial infection is
characterized by the presence of one or more of the following:
Methicillin resistant Staphylococci, Fluoroquinolone resistant
Staphylococci, Glycopepetide resistant Staphylococci,
Macrolide-Lincosamide-Streptogramin resistant Staphylococci, Linezolid
resistant Enterococci, Glycopepetide resistant Enterococci, β-lactam
resistant Enterococci, Penicillin resistant Streptococci, Macrolide
resistant Streptococci, Ketolide resistant Streptococci, Fluoroquinolone
resistant Streptococci, β-lactam resistant Haemophilus,
Fluoroquinolone resistant Haemophilus, Macrolide resistant Haemophilus,
Macrolide resistant Mycoplasma, Isoniazid resistant Mycobacterium,
Rifampin resistant Mycobacterium, or β-lactam resistant Moraxella,
comprising the step of adminstering to said patient a compound of formula
I:

[0018] The present invention also relates to methods for treating
resistant bacterial infections in patients. The present invention also
relates to treating resistant bacterial infections in patients using
compounds of the present invention in combination with one or more
additional antibacterial agents and/or one or more additional therapeutic
agents that increase the susceptibility of bacterial organisms to
antibiotics.

or a pharmaceutically acceptable salt thereof, wherein: [0037] V is
selected from nitrogen, CH, or CF; [0038] R3 is hydrogen or
C1-4 aliphatic, wherein: [0039] when R3 is C1-4
aliphatic, R3 is substituted with 0-3 groups independently selected
from OH, R5, or OR5; [0040] wherein: [0041] R5 is
C1-3 aliphatic, wherein: [0042] two R5 aliphatic groups may be
optionally taken together with the carbon to which they are bound to form
a C3-4 cycloalkyl ring; [0043] provided that if R3 is
hydrogen, then V is not nitrogen or CH; [0044] R4 is a C1-3
aliphatic group; and [0045] Ring C is a 6-membered heteroaryl ring having
1-2 nitrogens, wherein: [0046] Ring C is substituted with 1-3 groups
selected from R6; [0047] wherein: [0048] each R6 is
independently selected from OR7 or halogen; and [0049] R7 is
C1-4 aliphatic; or [0050] Ring C is an unsubstituted 2-pyrimidine
ring.

[0051] The present invention relates to a compound of formula I:

##STR00005##

or a pharmaceutically acceptable salt thereof, wherein: [0052] W is
selected from nitrogen, CH, or CF; [0053] X is selected from CH or CF;
[0054] Z is O or NH; [0055] R1 is phenyl or a 5-6 membered
heteroaryl ring having 1-3 heteroatoms independently selected from
oxygen, nitrogen, or sulfur, wherein: [0056] R1 is substituted with
0-3 groups independently selected from -(T)y-Ar, R', oxo, C(O)R',
CO2R', OR', N(R')2, SR', NO2, halogen, CN,
C(O)N(R')2, NR'C(O)R', SO2R', SO2N(R')2, or
NR'SO2R'; [0057] y is 0 or 1; [0058] T is a straight or branched
C1-4 alkylidene chain, wherein one methylene unit of T is optionally
replaced by --O--, --NH--, or --S--; [0059] each R' is independently
selected from hydrogen, C1-4 aliphatic, or a 5-6 membered saturated,
unsaturated, or aryl ring having 0-3 heteroatoms independently selected
from nitrogen, oxygen, or sulfur, wherein: [0060] R' is substituted with
0-3 groups independently selected from halogen, oxo, Ro,
N(RO)2, ORo, CO2Ro, NRoC(O)Ro,
C(O)N(RO)2, SO2Ro, SO2N(RO)2, or
NRoSO2Ro, wherein: [0061] each Ro is independently
selected from hydrogen, C1-4 aliphatic, or a 5-6 membered saturated,
unsaturated, or aryl ring having 0-3 heteroatoms independently selected
from nitrogen, oxygen, or sulfur, and wherein: [0062] two substituents on
adjacent positions of R1 may be taken together to form a 5-7
membered saturated, partially unsaturated, or aryl ring having 0-3
heteroatoms independently selected from nitrogen, oxygen, or sulfur;
[0063] Ar is a 3-8 membered saturated, unsaturated, or aryl ring, a 3-7
membered heterocyclic ring having 1-3 heteroatoms independently selected
from nitrogen, oxygen, or sulfur, or a 5-6 membered heteroaryl ring
having 1-3 heteroatoms independently selected from nitrogen, oxygen, or
sulfur, wherein: [0064] Ar is substituted with 0-3 groups independently
selected from R', oxo, CO2R', OR', N(R')2, SR', NO2,
halogen, CN, C(O)N(R')2, NR'C(O)R', SO2R', C(O)R',
SO2N(R')2, or NR'SO2R; [0065] R2 is selected from
hydrogen or a C1-3 aliphatic group; and [0066] Ring A is a 5-6
membered heteroaryl ring having 1-4 heteroatoms independently selected
from nitrogen, oxygen, or sulfur, provided that said ring has a
hydrogen-bond acceptor in the position adjacent to the point of
attachment to Ring B, wherein: [0067] Ring A is substituted with 0-3
groups independently selected from R', oxo, CO2R', OR', N(R')2,
SR', NO2, halogen, CN, C(O)N(R')2, NR'C(O)R', SO2R',
SO2N(R')2, or NR'SO2R', and wherein: [0068] two
substituents on adjacent positions of Ring A may be taken together to
form a 5-7 membered saturated, partially unsaturated, or aryl ring having
0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0069] The present invention also relates to a compound of formula VII:

##STR00006##

or a pharmaceutically acceptable salt thereof, wherein: [0070] V is
selected from nitrogen, CH, or CF; [0071] R3 is hydrogen or
C1-4 aliphatic, wherein: [0072] when R3 is C1-4
aliphatic, R3 is substituted with 0-3 groups independently selected
from OH, R5, or OR5; [0073] wherein: [0074] R5 is
C1-3 aliphatic, wherein: [0075] two R5 aliphatic groups may be
optionally taken together with the carbon to which they are bound to form
a C3-4 cycloalkyl ring; [0076] provided that if R3 is
hydrogen, then V is not nitrogen or CH; [0077] R4 is a C1-3
aliphatic group; and [0078] Ring C is a 6-membered heteroaryl ring having
1-2 nitrogens, wherein: [0079] Ring C is substituted with 1-3 groups
selected from R6; [0080] wherein: [0081] each R6 is
independently selected from OR7 or halogen; and [0082] R7 is
C1-4 aliphatic; or [0083] Ring C is an unsubstituted 2-pyrimidine
ring.

[0084] As used herein, the following definitions shall apply unless
otherwise indicated.

[0085] The phrase "optionally substituted" is used interchangeably with
the phrase "substituted or unsubstituted." Unless otherwise indicated, an
optionally substituted group may have a substituent at each substitutable
position of the group, and each substitution is independent of the other.

[0086] The term "aliphatic" or "aliphatic group", as used herein, means a
straight-chain or branched C1-C8 hydrocarbon chain that is
completely saturated or that contains one or more units of unsaturation,
or a monocyclic C3-C8 hydrocarbon or bicyclic C8-C12
hydrocarbon that is completely saturated or that contains one or more
units of unsaturation, but which is not aromatic (also referred to herein
as "carbocycle" or "cycloalkyl"), that has a single point of attachment
to the rest of the molecule wherein any individual ring in said bicyclic
ring system has 3-7 members. For example, suitable aliphatic groups
include, but are not limited to, linear or branched or alkyl, alkenyl,
alkynyl groups and hybrids thereof such as (cycloalkyl)alkyl,
(cycloalkenyl)alkyl or (cycloalkyl)alkenyl.

[0087] The terms "alkyl", "alkoxy", "hydroxyalkyl", "alkoxyalkyl", and
"alkoxycarbonyl", used alone or as part of a larger moiety include both
straight and branched chains containing one to twelve carbon atoms. The
terms "alkenyl" and "alkynyl" used alone or as part of a larger moiety
shall include both straight and branched chains containing two to twelve
carbon atoms.

[0088] The term "heteroatom" means nitrogen, oxygen, or sulfur and
includes any oxidized form of nitrogen and sulfur, and the quaternized
form of any basic nitrogen. Also the term "nitrogen" includes a
substitutable nitrogen of a heterocyclic ring. As an example, in a
saturated or partially unsaturated ring having 0-3 heteroatoms selected
from oxygen, sulfur or nitrogen, the nitrogen may be N (as in
3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR.sup.+ (as in
N-substituted pyrrolidinyl).

[0089] The term "unsaturated", as used herein, means that a moiety has one
or more units of unsaturation, and includes aryl rings.

[0090] The term "aryl" used alone or as part of a larger moiety as in
"aralkyl", "aralkoxy", or "aryloxyalkyl", refers to monocyclic, bicyclic
and tricyclic ring systems having a total of five to fourteen ring
members, wherein at least one ring in the system is aromatic and wherein
each ring in the system contains 3 to 7 ring members. The term "aryl" may
be used interchangeably with the term "aryl ring". The term "aryl" also
refers to heteroaryl ring systems as defined hereinbelow.

[0091] The term "heterocycle", "heterocyclyl", or "heterocyclic" as used
herein means non-aromatic, monocyclic, bicyclic or tricyclic ring systems
having five to fourteen ring members in which one or more ring members is
a heteroatom, wherein each ring in the system contains 3 to 7 ring
members.

[0092] The term "heteroaryl", used alone or as part of a larger moiety as
in "heteroaralkyl" or "heteroarylalkoxy", refers to monocyclic, bicyclic
and tricyclic ring systems having a total of five to fourteen ring
members, wherein at least one ring in the system is aromatic, at least
one ring in the system contains one or more heteroatoms, and wherein each
ring in the system contains 3 to 7 ring members. The term "heteroaryl"
may be used interchangeably with the term "heteroaryl ring" or the term
"heteroaromatic".

[0093] The term "hydrogen bond acceptor", as used herein, means an atom
capable of accepting a hydrogen bond. A typical hydrogen bond acceptor is
a sulfur, oxygen, or nitrogen atom, especially a nitrogen that is
sp2-hybridized, an ether oxygen, or a thioether sulfur. A preferred
hydrogen bond acceptor is a nitrogen that is sp2-hybridized.

[0094] A combination of substituents or variables is permissible only if
such a combination results in a stable or chemically feasible compound. A
stable compound or chemically feasible compound is one that is not
substantially altered when kept at a temperature of 40° C. or
less, in the absence of moisture or other chemically reactive conditions,
for at least a week.

[0095] It will be apparent to one skilled in the art that certain
compounds of this invention may exist in tautomeric forms, all such
tautomeric forms of the compounds being within the scope of the
invention.

[0096] Unless otherwise stated, structures depicted herein are also meant
to include all stereochemical forms of the structure; i.e., the R and S
configurations for each asymmetric center. Therefore, single
stereochemical isomers as well as enantiomeric and diastereomeric
mixtures of the present compounds are within the scope of the invention.
Unless otherwise stated, structures depicted herein are also meant to
include compounds that differ only in the presence of one or more
isotopically enriched atoms. For example, compounds having the present
structures except for the replacement of a hydrogen by a deuterium or
tritium, or the replacement of a carbon by a 13C-- or
14C-enriched carbon are within the scope of this invention. Such
compounds are useful, for example, as analytical tools or probes in
biological assays.

[0097] Examples of suitable Ring A moieties are set forth in Table 1
below.

[0098] According to one embodiment, Ring A of formula I is a 5-membered
heteroaryl ring having 1-4 heteroatoms independently selected from
nitrogen, oxygen, or sulfur, provided that said ring has a hydrogen-bond
acceptor in the position adjacent to the point of attachment to Ring B,
wherein said Ring A is optionally substituted as defined herein supra.

[0099] According to another embodiment, Ring A of formula I is a
6-membered heteroaryl ring having 1-3 nitrogens, provided that said ring
has a nitrogen atom in the position adjacent to the point of attachment
to Ring B, wherein said Ring A is optionally substituted as defined
herein supra.

[0101] In other embodiments, the Ring A moieties of formula I are selected
from rings a, f, l, s, w, y, and z, wherein each Ring A is optionally
substituted as defined above.

[0102] When Ring A of formula I is a bicyclic heteroaryl ring, preferred
bicyclic Ring A moieties include benzothiazole, benzimidazole,
benzoxazole, and quinoline.

[0103] According to one embodiment, substituents on Ring A of formula I,
if present, are selected from oxo, N(R')2, C(O)N(R')2,
CO2R', halogen, N(R')SO2R', C(O)R', OR', or R'. According to
another embodiment, R' substituents on Ring A of formula I include
methyl, ethyl, propyl, piperazinyl, piperidinyl, or morpholinyl, wherein
said R' groups are optionally substituted with Ro, N(RO)2
or ORo.

[0104] According to one embodiment, the R1 group of formula I is
optionally substituted phenyl.

[0105] According to another embodiment, the R1 group of formula I is
an optionally substituted 5-membered heteroaryl ring having 1-3
heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0106] According to another embodiment, the R1 group of formula I is
an optionally substituted 5-membered heteroaryl ring having 1-3
nitrogens.

[0107] Yet another embodiment of the present invention relates to a
compound of formula I wherein R1 is an optionally substituted
6-membered heteroaryl ring having 1-2 nitrogens.

[0108] In certain embodiments, the R1 group of formula I is selected
from an optionally substituted phenyl or 5-6 membered heteroaryl ring
having 1-2 nitrogens. In other embodiments, the R1 group of formula
I is selected from an optionally substituted pyrid-2-yl, pyrid-3-yl,
pyrid-4-yl, pyridone, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl,
pyrimidin-6-yl, imidazol-1-yl, imidazol-2-yl, imidazol-4-yl, or
imidazol-5-yl ring. According to yet another embodiment, the R1
group of formula I is an optionally substituted ring selected from
pyrid-3-yl, pyrid-4-yl, pyridone, pyrimidin-5-yl, or imidazol-1-yl.

[0110] According to another embodiment, substituents on the R1 group
of formula I, when present, include an optionally substituted C1-3
alkyl group.

[0111] According to another embodiment, substituents on the R1 group
of formula I, when present, include a C1-3 alkyl group optionally
substituted with an OH group.

[0112] According to another embodiment, substituents on the R1 group
of formula I, when present, include a C1-3 alkyl group substituted
with an OH group.

[0113] According to another embodiment, substituents on the C1-3
alkyl group, when present, include methyl, gem-dimethyl, ethyl, propyl,
isopropyl, cyclopropyl, or cyclobutyl, and an OH group.

[0114] According to one embodiment, R1 is substituted with
-(T)y-Ar wherein T is a straight or branched C1-3 alkylidene
chain wherein one methylene unit of T is optionally replaced by --O--,
--NH--, or --S--. According to another embodiment, T is a straight or
branched C1-3 alkylidene chain wherein one methylene unit of T is
replaced by --O--, --NH--, or --S--. Yet another embodiment of the
present invention relates to a compound of formula I wherein R1 is
substituted with -(T)y-Ar and Ar is an optionally substituted 5-6
membered saturated ring having 1-2 heteroatoms independently selected
from oxygen, nitrogen, or sulfur. According to another embodiment, the Ar
group of formula I is an optionally substituted 5-membered heteroaryl
ring having 1-3 heteroatoms independently selected from nitrogen, oxygen,
or sulfur. According to yet another embodiment, the Ar group of formula I
is an optionally substituted 6-membered heteroaryl ring having 1-3
nitrogens. Yet another embodiment relates to a compound of formula I
wherein Ar is optionally substituted phenyl.

[0115] When the R1 group of formula I is substituted with
-(T)y-Ar, examples of substituents on Ar include halogen, OR', R',
CO2R', SO2R', oxo, and C(O)R'.

[0116] According to one embodiment, when two substituents on adjacent
positions of R1 of formula I are taken together to form an
optionally substituted ring fused to R1, rings formed thereby
include 5-6 membered saturated, partially unsaturated, or aryl rings
having 0-2 heteroatoms independently selected from nitrogen, oxygen, or
sulfur. According to another embodiment, said ring fused to R1 is
selected from a 5-membered saturated ring having two oxygens or a
6-membered saturated ring having two oxygens. Examples of substituents on
said ring fused to R1 include halogen, such as fluorine.

[0117] One embodiment of the present invention relates to a compound of
formula I wherein R2 is selected from methyl, ethyl, isopropyl, or
cyclopropyl. According to another embodiment, R2 is methyl or ethyl.
According to yet another embodiment, R2 of formula I is ethyl.

[0118] According to one embodiment, the present invention relates to a
compound of formula I wherein Z is NH.

[0119] According to another embodiment, the present invention relates to a
compound of formula I wherein Z is O.

[0120] Compounds of the present invention fall within the genus of
compounds described in PCT/US01/48855. However, applicants have
discovered that the presence of the Ring A moiety, as defined above,
imparts surprising and unexpectedly increased gyrase inhibitory, Topoly
activity, and antimicrobial potency.

[0121] According to one embodiment, the present invention relates to a
compound of formula II:

##STR00041##

or a pharmaceutically acceptable salt thereof, wherein Z, R2 and
Ring A are as defined above and the imidazole ring depicted is optionally
substituted in the 4-position with C(O)N(R')2 and/or substituted in
the 2-position with R'.

[0122] According to one embodiment, the imidazole ring of formula II is
optionally substituted with a C1-3 alkyl group.

[0123] According to another embodiment, the imidazole ring of formula II
is substituted with a C1-3 alkyl group that is optionally
substituted with an OH group.

[0124] According to another embodiment, the imidazole ring of formula II
is substituted with a C1-3 alkyl group that is substituted with an
OH group.

[0125] According to another embodiment, the imidazole ring of formula II
is substitued with a methyl, gem-dimethyl, ethyl, propyl, isopropyl,
cyclopropyl, or cyclobutyl, and an OH group.

[0126] According to another embodiment, the present invention relates to a
compound of formula II-a:

##STR00042##

or a pharmaceutically acceptable salt thereof, wherein Z, R2, R',
and Ring A are as defined above.

[0127] Other embodiments describing R2 and Ring A groups of formula
II-a are those described for formula I above.

[0128] Other embodiments describing R' groups of formula II-a are selected
from hydrogen or C1-4 aliphatic.

[0129] According to one embodiment, the present invention relates to a
compound of formula II or II-a wherein Z is NH.

[0130] According to another embodiment, the present invention relates to a
compound of formula II or II-a wherein Z is O.

[0131] According to another embodiment, the present invention relates to a
compound of formula III:

[0132] Other embodiments describing R2 and Ring A groups of formula
III are those described for formula I above.

[0133] Other embodiments describing substituents on the pyridone ring of
formula III are those described above as preferred substituents on
R1 of formula I.

[0134] According to another embodiment, the pyridone ring of formula III
is optionally substituted with a C1-3 alkyl group.

[0135] According to another embodiment, the pyridone ring of formula III
is substituted with a C1-3 alkyl group that is optionally
substituted with an OH group.

[0136] According to another embodiment, the pyridone ring of formula III
is substituted with a C1-3 alkyl group that is substituted with an
OH group.

[0137] According to another embodiment, the pyridone ring of formula III
is substitued with a methyl, gem-dimethyl, ethyl, propyl, isopropyl,
cyclopropyl, or cyclobutyl, and an OH group.

[0138] According to one embodiment, the present invention relates to a
compound of formula III wherein Z is NH.

[0139] According to another embodiment, the present invention relates to a
compound of formula III wherein Z is O.

[0140] According to another embodiment, the present invention relates to a
compound of formula III-a:

##STR00044##

or a pharmaceutically acceptable salt thereof, wherein Z, R', R2 and
Ring A are as defined above.

[0141] Other embodiments describing R2 groups of formula III-a are
those described for R2 groups of formula I above.

[0142] Other embodiments describing Ring A groups of formula III-a are
those described for Ring A groups of formula I above.

[0143] In certain embodiments, the R' substituents on the pyridone ring of
formula III-a are selected from hydrogen or C1-4 aliphatic wherein
R' is optionally substituted with phenyl or pyridyl. In other
embodiments, the R' substituents on the pyridone ring of formula III-a
are selected from methyl, ethyl, t-butyl, isobutyl, cyclopropyl,
isopropyl, CH2-phenyl, CH2pyridin-3-yl, CH2piperidinyl,
CH2cyclopropyl, or CH2CH2OCH3.

[0144] According to another embodiment, the pyridone ring of formula III-a
is optionally substituted with a C1-3 alkyl group.

[0145] According to another embodiment, the pyridone ring of formula III-a
is substituted with a C1-3 alkyl group that is optionally
substituted with an OH group.

[0146] According to another embodiment, the pyridone ring of formula III-a
is substituted with a C1-3 alkyl group that is substituted with an
OH group.

[0147] According to another embodiment, the pyridone ring of formula III-a
is substitued with a methyl, ethyl, propyl, isopropyl, cyclopropyl, or
cyclobutyl, and an OH group.

[0148] According to one embodiment, the present invention relates to a
compound of formula III-a wherein Z is NH.

[0149] According to another embodiment, the present invention relates to a
compound of formula III-a wherein Z is O.

[0150] Yet another embodiment of the present invention relates to a
compound of formula IV:

##STR00045##

or a pharmaceutically acceptable salt thereof, wherein y, Z, T, Ar,
R2 and Ring A are as defined above.

[0151] According to one embodiment, the pyridone ring of formula IV is
optionally substituted with a C1-3 alkyl group.

[0152] According to another embodiment, the pyridone ring of formula IV is
substituted with a C1-3 alkyl group that is optionally substituted
with an OH group.

[0153] According to another embodiment, the pyridone ring of formula IV is
substituted with a C1-3 alkyl group that is substituted with an OH
group.

[0154] According to another embodiment, the pyridone ring of formula IV is
substitued with a methyl, gem-dimethyl, ethyl, propyl, isopropyl,
cyclopropyl, or cyclobutyl, and an OH group.

[0155] Other embodiments describing Ring A and R2 groups of formula
IV are those set forth for those Ring A and R2 groups of formula I,
supra.

[0156] According to one embodiment, the Ar group of formula IV is an
optionally substituted 5-6 membered saturated ring having 1-2 heteroatoms
independently selected from oxygen, nitrogen, or sulfur.

[0157] According to another embodiment, the Ar group of formula IV is an
optionally substituted 5-membered heteroaryl ring having 1-3 heteroatoms
independently selected from nitrogen, oxygen, or sulfur.

[0158] According to another embodiment, the Ar group of formula IV is an
optionally substituted 6-membered heteroaryl ring having 1-3 nitrogens.

[0173] Other embodiments describing the R2 group of formula VI are
those set forth for the R2 group of formula I, supra.

[0174] According to one embodiment, the Ar group of formula VI is an
optionally substituted 5-6 membered saturated ring having 1-2 heteroatoms
independently selected from oxygen, nitrogen, or sulfur.

[0175] According to another embodiment, the Ar group of formula VI is an
optionally substituted 5-membered heteroaryl ring having 1-3 heteroatoms
independently selected from nitrogen, oxygen, or sulfur.

[0176] According to another embodiment, the Ar group of formula VI is an
optionally substituted 6-membered heteroaryl ring having 1-3 nitrogens.

[0178] According to another embodiment, the pyridone ring of formula VI is
optionally substituted with a C1-3 alkyl group.

[0179] According to another embodiment, the pyridone ring of formula VI is
substituted with a C1-3 alkyl group that is optionally substituted
with an OH group.

[0180] According to another embodiment, the pyridone ring of formula VI is
substituted with a C1-3 alkyl group that is substituted with an OH
group.

[0181] According to another embodiment, the pyridone ring of formula VI is
substitued with a methyl, gem-dimethyl, ethyl, propyl, isopropyl,
cyclopropyl, or cyclobutyl, and an OH group.

[0182] According to one embodiment, the present invention relates to a
compound of formula VI wherein Z is NH.

[0183] Examples of substituents on the Ar group of formula VI include
halogen, OR', R', CO2R', SO2R', oxo, and C(O)R'.

[0184] According to another embodiment, the present invention relates to a
compound of formula VI wherein Z is O.

[0185] According to another embodiment, the present invention relates to a
compound of formula VII:

##STR00048##

or a pharmaceutically acceptable salt thereof, wherein: [0186] V is
selected from nitrogen, CH, or CF; [0187] R3 is hydrogen or
C1-4 aliphatic, wherein: [0188] when R3 is C1-4 aliphatic,
R3 is substituted with 0-3 groups independently selected from OH,
R5, or OR5; [0189] wherein: [0190] R5 is C1-3
aliphatic, wherein: [0191] two R5 aliphatic groups may be optionally
taken together with the carbon to which they are bound to form a
C3-4 cycloalkyl ring; [0192] provided that if R3 is hydrogen,
then V is not nitrogen or CH; [0193] R4 is a C1-3 aliphatic
group; and [0194] Ring C is a 6-membered heteroaryl ring having 1-2
nitrogens, wherein: [0195] Ring C is substituted with 1-3 groups
selected from R6; [0196] wherein: [0197] each R6 is
independently selected from OR7 or halogen; and [0198] R7 is
C1-4 aliphatic; or [0199] Ring C is an unsubstituted 2-pyrimidine
ring.

[0200] According to another embodiment, the present invention relates to a
compound of formula VII:

##STR00049##

or a pharmaceutically acceptable salt thereof, wherein: [0201] V is
selected from nitrogen or CH; [0202] R3 is C1-4 aliphatic,
wherein: [0203] when R3 is C1-4 aliphatic, R3 is
substituted with 0-3 groups independently selected from OH, R5, or
OR5; [0204] wherein: [0205] R5 is C1-3 aliphatic,
wherein: [0206] two R5 aliphatic groups may be optionally taken
together with the carbon to which they are bound to form a C3-4
cycloalkyl ring; [0207] R4 is a C1-3 aliphatic group; and
[0208] Ring C is a 6-membered heteroaryl ring having 1-2 nitrogens,
wherein: [0209] Ring C is substituted with 1-3 groups selected from
R6; [0210] wherein: [0211] each R6 is independently selected
from OR7 or halogen; and [0212] R7 is C1-4 aliphatic.

[0213] According to another embodiment of compounds of formula VII, Ring C
is a 2-pyridyl ring substituted with one occurrence of R6.

[0214] According to another embodiment of compounds of formula VII, Ring C
is

##STR00050##

[0215] According to another embodiment of compounds of formula VII,
R6 is selected from halogen or OR7.

[0216] According to another embodiment of compounds of formula VII,
R7 is methyl.

[0217] According to another embodiment of compounds of formula VII,
R6 is fluoro.

[0218] According to another embodiment of compounds of formula VII, the

##STR00051##

radical is:

##STR00052##

[0219] According to another embodiment of compounds of formula VII,
R3 is an optionally substituted C1-3 alkyl group.

[0220] According to another embodiment of compounds of formula VII,
R3 is a C1-3 alkyl group optionally substituted with an OH
group.

[0221] According to another embodiment of compounds of formula VII,
R3 is a C1-3 alkyl group substituted with an OH group

[0222] According to another embodiment of compounds of formula VII, V is
nitrogen and R3 is substituted with two groups independently
selected from R5 and OH.

[0223] According to another embodiment of compounds of formula VII, V is
CH and R3 is substituted with two groups independently selected from
R5 and OH.

[0224] According to another embodiment of compounds of formula VII,
R3 is substituted with one R5 group and one OH group.

[0225] According to another embodiment of compounds of formula VII,
R3 is a C1-3 alkyl group substituted with methyl, gem-dimethyl,
ethyl, propyl, isopropyl, cyclopropyl, or cyclobutyl, and an OH group.

[0226] According to another embodiment, the present invention relates to a
compound of formula VIIA, VIIB or VIIC:

or a pharmaceutically acceptable salt thereof, wherein: [0267] V is
selected from nitrogen, CH, or CF; [0268] R3 is hydrogen or
C1-4 aliphatic, wherein: [0269] when R3 is C1-4 aliphatic,
R3 is substituted with 0-3 groups independently selected from OH,
R5, or OR5; [0270] wherein: [0271] R5 is C1-3
aliphatic, wherein: [0272] two R5 aliphatic groups may be optionally
taken together with the carbon to which they are bound to form a
C3-4 cycloalkyl ring; [0273] provided that if R3 is hydrogen,
then V is not nitrogen or CH; [0274] R4 is a C1-3 aliphatic
group; and [0275] Ring C is a 6-membered heteroaryl ring having 1-2
nitrogens, wherein: [0276] Ring C is substituted with 1-3 groups
selected from R6; [0277] wherein: [0278] each R6 is
independently selected from OR7 or halogen; and [0279] R7 is
C1-4 aliphatic; or [0280] Ring C is an unsubstituted
2-pyrimidine ring.

[0281] According to another embodiment, the method of the present
invention comprises the step of adminstering to said patient a compound
of formula VIIA, VIIB, or VIIC:

[0291] According to another embodiment of the methods of the present
invention, the Methicillin resistant Staphylococcci are selected from
Methicillin resistant Staphylococcus aureus, Methicillin resistant
Staphylococcus epidermidis, or Methicillin resistent Coagulase negative
staphylcoccus.

[0292] According to another embodiment of the methods of the present
invention, the Fluoroquinolone resistant Staphylococci are selected from
Fluoroquinolone resistant Staphylococcus aureus, Fluoroquinolone
resistant Staphylococcus epidermidis, or Fluoroquinolone resistant
Coagulase negative staphylcoccus.

[0294] According to another embodiment of the methods of the present
invention, the Macrolide-Lincosamide-Streptogramin resistant
Staphylococci is Macrolide-Lincosamide-Streptogramin resistant
Staphylococcus aureus.

[0295] According to another embodiment of the methods of the present
invention, the Linezolid resistant Enterococci are selected from
Linezolid resistant Enterococcus faecalis, or Linezolid resistant
Enterococcus faecium.

[0296] According to another embodiment of the methods of the present
invention, the Glycopepetide resistant Enterococci are selected from
Vancomycin resistant Enterococcus faecium or Vancomycin resistant
Enterococcus faecalis.

[0297] According to another embodiment of the methods of the present
invention, the β-lactam resistant Enterococcus faecalis and
β-lactam resistant Enterococcus faecium.

[0298] According to another embodiment of the methods of the present
invention, the Penicillin resistant Streptococci include Penicillin
resistant Streptococcus pneumoniae.

[0299] According to another embodiment of the methods of the present
invention, the Macrolide resistant Streptococci is Macrolide resistant
Streptococcus pneumonia.

[0300] According to another embodiment of the methods of the present
invention, the Ketolide resistant Streptococci are selected from
Macrolide resistant Streptococcus pneumoniae and Ketolide resistant
Streptococcus pyogenes.

[0301] According to another embodiment of the methods of the present
invention, the Fluoroquinolone resistant Streptococci is Fluoroquinolone
resistant Streptococcus pneumoniae.

[0302] According to another embodiment of the methods of the present
invention, the β-lactam resistant Haemophilus is β-lactam
resistant Haemophilus influenzae.

[0303] According to another embodiment of the methods of the present
invention, the Fluoroquinolone resistant Haemophilus is Fluoroquinolone
resistant Haemophilus influenzae.

[0304] According to another embodiment of the methods of the present
invention, the Macrolide resistant Haemophilus is Macrolide resistant
Haemophilus influenzae.

[0305] According to another embodiment of the methods of the present
invention, the Macrolide resistant Mycoplasma is Macrolide resistant
Mycoplama pneumoniae.

[0306] According to another embodiment of the methods of the present
invention, the Isoniazid resistant Mycobacterium is Isoniazid resistant
Mycobacterium tuberculosis.

[0307] According to another embodiment of the methods of the present
invention, the Rifampin resistant Mycobacterium is Rifampin resistant
Mycobacterium tuberculosis.

[0308] According to another embodiment of the methods of the present
invention, the β-lactam resistant Moraxella is β-lactam
resistant Moraxella catarrhalis.

[0312] According to another embodiment of the present invention, the
methods further comprise the step of administering to the patient one or
more additional therapeutic antibacterial agents other than a compound of
the present invention (see, e.g. http://www.fda.gov/cvm).

[0313] According to another embodiment of the present invention, the
methods further comprise the step of administering to said patient one or
more additional therapeutic agents either as part of a multiple dosage
form together with said compound or as a separate dosage form wherein
said one or more additional therapeutic agents include an antibiotic
selected from a natural penicillin, a penicillinase-resistant penicillin,
an antipseudomonal penicillin, an aminopenicillin, a first generation
cephlosporin, a second generation cephalosporin, a third generation
cephalosporin, a fourth generation cephalosporin, a carbapenem, a
cephamycin, a monobactam, a quinolone, a fluoroquinolone, an
aminoglycoside, a macrolide, a ketolide, a tetracycline, a glycopeptide,
a streptogramin, an oxazolidone, a rifamycin, or other antibiotics.

[0314] According to another embodiment of the present invention, the
methods further comprise the step of administering to said human one or
more additional therapeutic agents either as part of a multiple dosage
form together with said compound or as a separate dosage form wherein
said one or more additional therapeutic agents include an antibiotic
selected from a natural penicillin, a penicillinase-resistant penicillin,
an antipseudomonal penicillin, an aminopenicillin, a first generation
cephlosporin, a second generation cephalosporin, a third generation
cephalosporin, a fourth generation cephalosporin, a carbapenem, a
cephamycin, a monobactam, a quinolone, a fluoroquinolone, an
aminoglycoside, a macrolide, a ketolide, a tetracycline, a glycopeptide,
a streptogramin, an oxazolidone, a rifamycin, or other antibiotics.

[0315] According to another embodiment of the present invention, the
methods further comprise the step of administering to said patient one or
more additional therapeutic agents either as part of a multiple dosage
form together with said compound or as a separate dosage form wherein
said one or more additional therapeutic agents are selected from a
natural penicillin including Benzathine penicillin G, Penicillin G and
Penicillin V, from a penicillinase-resistant penicillin including
Cloxacillin, Dicloxacillin, Nafcillin and Oxacillin, from a
antipseudomonal penicillin including Carbenicillin, Mezlocillin,
Pipercillin, Pipercillin/tazobactam, Ticaricillin and
Ticaricillin/Clavulanate, from an aminopenicillin including Amoxicillin,
Ampicillin and Ampicillin/Sulbactam, from a first generation
cephalosporin including Cefazolin, Cefadroxil, Cephalexin and Cephadrine,
from a second generation cephalosporin including Cefaclor, Cefaclor-CD,
Cefamandole, Cefonacid, Cefprozil, Loracarbef and Cefuroxime, from a
third generation cephalosporin including Cefdinir, Cefixime,
Cefoperazone, Cefotaxime, Cefpodoxime, Ceftazidime, Ceftibuten,
Ceftizoxme and Ceftriaxone, from a fourth generation cephalosporin
including Cefepime, from a Cephamycin including Cefotetan and Cefoxitin,
from a carbapenem including Imipenem and Meropenem, from a monobactam
including Aztreonam, from a quinolone including Cinoxacin, Nalidixic
acid, Oxolininc acid and Pipemidic acid, from a fluoroquinolone including
Cirpofloxacin, Enoxacin, Gatifloxacin, Grepafloxacin, Levofloxacin,
Lomefloxacin, Moxifloxacin, Norfloxacin, Ofloxacin and Sparfloxacin, from
an aminoglycoside including Amikacin, Gentamicin, Kanamycin, Neomycin,
Netilmicin, Spectinomycin, Streptomycin and Tobramycin, from a macrolide
including Azithromycin, Clarithromycin and Erythromycin, from a ketolide
including Telithromycin, from a Tetracycline including Chlortetracycline,
Demeclocycline, Doxycycline, Minocycline and Tetracycline, from a
glycopeptide including Oritavancin, Teicoplanin and Vancomycin, from a
streptogramin including Dalfopristin/quinupristin, from an oxazolidone
including Linezolid, from a Rifamycin including Rifabutin and Rifampin
and from other antibiotics including bactitracin, chloramphenicol,
clindamycin, isoniazid, metronidazole, polymyxin B, pyrazinamide, and
trimethoprim/sulfamethoxazole.

[0316] According to another embodiment of the present invention, the
methods further comprise the step of administering to said human one or
more additional therapeutic agents either as part of a multiple dosage
form together with said compound or as a separate dosage form wherein
said one or more additional therapeutic agents are selected from a
natural penicillin including Benzathine penicillin G, Penicillin G and
Penicillin V, from a penicillinase-resistant penicillin including
Cloxacillin, Dicloxacillin, Nafcillin and Oxacillin, from a
antipseudomonal penicillin including Carbenicillin, Mezlocillin,
Pipercillin, Pipercillin/tazobactam, Ticaricillin and
Ticaricillin/Clavulanate, from an aminopenicillin including Amoxicillin,
Ampicillin and Ampicillin/Sulbactam, from a first generation
cephalosporin including Cefazolin, Cefadroxil, Cephalexin and Cephadrine,
from a second generation cephalosporin including Cefaclor, Cefaclor-CD,
Cefamandole, Cefonacid, Cefprozil, Loracarbef and Cefuroxime, from a
third generation cephalosporin including Cefdinir, Cefixime,
Cefoperazone, Cefotaxime, Cefpodoxime, Ceftazidime, Ceftibuten,
Ceftizoxme and Ceftriaxone, from a fourth generation cephalosporin
including Cefepime, from a Cephamycin including Cefotetan and Cefoxitin,
from a carbapenem including Imipenem and Meropenem, from a monobactam
including Aztreonam, from a quinolone including Cinoxacin, Nalidixic
acid, Oxolininc acid and Pipemidic acid, from a fluoroquinolone including
Cirpofloxacin, Enoxacin, Gatifloxacin, Grepafloxacin, Levofloxacin,
Lomefloxacin, Moxifloxacin, Norfloxacin, Ofloxacin and Sparfloxacin, from
an aminoglycoside including Amikacin, Gentamicin, Kanamycin, Neomycin,
Netilmicin, Spectinomycin, Streptomycin and Tobramycin, from a macrolide
including Azithromycin, Clarithromycin and Erythromycin, from a ketolide
including Telithromycin, from a Tetracycline including Chlortetracycline,
Demeclocycline, Doxycycline, Minocycline and Tetracycline, from a
glycopeptide including Oritavancin, Teicoplanin and Vancomycin, from a
streptogramin including Dalfopristin/quinupristin, from an oxazolidone
including Linezolid, from a Rifamycin including Rifabutin and Rifampin
and from other antibiotics including bactitracin, chloramphenicol,
clindamycin, isoniazid, metronidazole, polymyxin B, pyrazinamide, and
trimethoprim/sulfamethoxazole.

[0317] According to another embodiment of the present invention, the
methods further comprise the step of administering to said patient one or
more additional therapeutic agents either as part of a multiple dosage
form together with said compound or as a separate dosage form wherein
said one or more additional therapeutic agents are selected from a
natural penicillin including Penicillin G, from a penicillinase-resistant
penicillin including Nafcillin and Oxacillin, from an antipseudomonal
penicillin including Pipercillin/tazobactam, from an aminopenicillin
including Amoxicillin, from a first generation cephalosporin including
Cephalexin, from a second generation cephalosporin including Cefaclor,
Cefaclor-CD and Cefuroxime, from a third generation cephalosporin
including Ceftazidime and Ceftriaxone, from a fourth generation
cephalosporin including Cefepime, from a fluoroquinolone including
Cirpofloxacin, Gatifloxacin, Levofloxacin and Moxifloxacin, from an
aminoglycoside including Tobramycin, from a macrolide including
Azithromycin and Clarithromycin, from a Tetracycline including
Doxycycline, from a glycopeptide including Vancomycin, from a Rifamycin
including Rifampin and from other antibiotics including isoniazid,
pyrazinamide, or trimethoprim/sulfamethoxazole.

[0318] According to another embodiment of the present invention, the
methods further comprise the step of administering to said human one or
more additional therapeutic agents either as part of a multiple dosage
form together with said compound or as a separate dosage form wherein
said one or more additional therapeutic agents are selected from a
natural penicillin including Penicillin G, from a penicillinase-resistant
penicillin including Nafcillin and Oxacillin, from an antipseudomonal
penicillin including Pipercillin/tazobactam, from an aminopenicillin
including Amoxicillin, from a first generation cephalosporin including
Cephalexin, from a second generation cephalosporin including Cefaclor,
Cefaclor-CD and Cefuroxime, from a third generation cephalosporin
including Ceftazidime and Ceftriaxone, from a fourth generation
cephalosporin including Cefepime, from a fluoroquinolone including
Cirpofloxacin, Gatifloxacin, Levofloxacin and Moxifloxacin, from an
aminoglycoside including Tobramycin, from a macrolide including
Azithromycin and Clarithromycin, from a Tetracycline including
Doxycycline, from a glycopeptide including Vancomycin, from a Rifamycin
including Rifampin and from other antibiotics including isoniazid,
pyrazinamide, or trimethoprim/sulfamethoxazole.

[0319] According to another embodiment of the present invention, the
methods further comprise the step of administering to a patient, one or
more additional therapeutic agents that increase the susceptibility of
bacterial organisms to antibiotics.

[0320] According to another embodiment of the present invention, the
methods further comprise the step of administering to a human, one or
more additional therapeutic agents that increase the susceptibility of
bacterial organisms to antibiotics.

[0321] According to another embodiment of the present invention, the
methods further comprise the step of administering to a patient, one or
more additional therapeutic agents that increase the susceptibility of
bacterial organisms to antibiotics including a biofilm inhibitor.

[0322] According to another embodiment of the present invention, the
methods further comprise the step of administering to a human, one or
more additional therapeutic agents that increase the susceptibility of
bacterial organisms to antibiotics including a biofilm inhibitor.

[0323] Exemplary structures of formula I are set forth in Table 2 below:

[0326] The compounds of this invention may be prepared in general by
methods known to those skilled in the art for analogous compounds, as
illustrated by the general Schemes I, II, III, IV, and V shown below and
the Examples set forth infra.

##STR00557##

[0327] Scheme I above shows a general method for preparing
N'-alkyl-N-cyanoureas 3 useful in the preparation of the compounds of the
present invention wherein Z is NH. At step (a), cyanamide 2 was treated
with an alkyl isocyanate in aqueous sodium hydroxide to afford, after
acidification, compound 3. One of skill in the art would recognize that a
variety of alkyl isocyanates would be amenable to the reaction conditions
of Scheme Ito form a variety of N'-alkyl-N-cyanoureas.

##STR00558##

[0328] Scheme II above shows a general method for preparing the
benzimidazole compounds of the present invention wherein Z is NH or O.
The bromo-aniline 4 was treated with sodium perborate and acetic acid to
form the difluoro-nitro compound 5. Compound 5 was treated with Ring A in
the presence of sodium hydride to afford the bi-aryl compound 6. The
remaining fluoro group of compound 6 was displaced with ammonia to form
the amino compound 7. The 2-nitro-5-bromoaniline 7 was then coupled to an
aryl boronic acid, at step (d), in the presence of palladium to form the
tri-aryl compound 8. The nitro group of compound 8 was reduced to form a
diamino compound which was either treated with an N'-alkyl-N-cyanourea 3
or with an N,N-dialkylureamido-2-methyl-2-thiopseudourea 3a to form a
benzimidazole compound of formula I wherein Z is NH 9.

[0329] Alternatively, intermediate 8 may be used to form compounds of
formula I wherein Z is 0. Compound 10 was formed by treating 8, after
reduction to the diamino compound, with 2-methyl-2-thiopseudourea and
R2-chloroformate according to the method described by L. I. Kruse et
al, J. Med. Chem. 1989, 32, 409-417. One of ordinary skill in the art
would recognize that the reactions depicted in Scheme II above are
amenable to a variety of R1 and Ring A groups of the present
invention.

[0330] In an alternative method, intermediate 8 was treated with either
N,N-diethlycarboxy-2-methyl-2-thiopseudourea or
N,N-diethlyureamido-2-methyl-2-thiopseudourea to form compounds 10 and 9,
respectively. The syntheses of both
N,N-diethlycarboxy-2-methyl-2-thiopseudourea and
N,N-diethlyureamido-2-methyl-2-thiopseudourea are described in the
Examples set forth infra.

##STR00559##

[0331] Scheme III above shows a general method for preparing compounds of
formula II-a using methods substantially similar to those described by
Kiyomori, A.; Marcoux, J.-F.; Buchwald, S. L., Tetrahedron Letters, vol.
40, (1999) 2657-2660. Compound 7 was treated with diboronic ester in the
presence of Pd(dppf)/potassium acetate in DMSO at 80° C. to afford
intermediate 11. Compound II was treated with 4-C(O)N(R')-2-imidazole in
the presence of copper acetate to form the 4-C(O)N(R')-2-imidazol-1-yl
compound 12. Compounds of formula II-a were prepared from compound 12 as
described in Scheme II, steps (e), (f), and (g).

[0332] Although 4-C(O)N(R')-2-imidazole was used to exemplify, one of
ordinary skill in the art would recognize that a variety of R1
groups are amenable to the displacement reaction at step (c) to form a
variety of compounds of the present invention. Generally, the boronate
intermediate 11 may be treated with a variety of R1-halides or
R1-triflates, using methods well known to one of ordinary skill in
the art, to form intermediate compounds 12' as shown below. Using the
methods recited herein and those known to one of ordinary skill in the
art, compounds 12' are useful for preparing compounds 9 and 10 of the
present invention as depicted above at Scheme II.

##STR00560##

##STR00561##

[0333] Scheme IV above shows an alternate method for preparing compounds
of formula II-a. Compound 13 was nitrated to form 14. Compound 14 was
treated with ammonium hydroxide to form the amino compound 15. The bromo
group of compound 15 was treated with the BrZn-Ring A reagent in the
presence of Pd(PPh3)4 in THF to form compound 16. Compound 16
was treated with the 4-C(O)N(R')-2-imidazole in the presence of sodium
carbonate to form the 4-C(O)N(R')-2-imidazol-1-yl compound 18. Compounds
of formula II-a were then prepared from compound 18 as described in
Scheme II, steps (e), (f), and (g).

##STR00562## ##STR00563##

[0334] Scheme V above shows a general method for preparing compounds of
formula VII. Compound 19 (purchased commercially from CB Research)
underwent Suzuki-type coupling with commercial reagents (purchased from
Aldrich or Manchester Organics Limited) of type 20 to form intermediate
21. Compound 21 was brominated with bromine in acetic acid to form
bromide 22, then nitrated with nitric acid in the presence of TFA and
TFAA to form intermediate 23. Acidic hydrolysis of compound 23 provided
aniline 24. The bromo group of compound 24 was converted into boronate
ester 25 in the presence of Pd(PPh3)4 in buffered dioxane.
Compound 25 underwent Suzuki-type coupling with intermediate 26 (prepared
from the di-bromoheteroaryl ring D [commercially available from Aldrich,
etc.] according to the procedure of Wang, X. et al., Tetrahedron Letters,
vol. 41, (2000) 4335-4338) to prepare nitro aniline intermediate 27.
Raney-nickel reduction of the nitro group in compound 27, followed by
treatment with N,N-dialkylureamido-2-methyl-2-thiopseudourea 3a afforded
benzimidazole compounds of formula VII. Compounds of formula VII wherein
Ring C is:

##STR00564##

and R6 is OMe are either prepared from compounds of formula 20,
using scheme V, wherein formula 20 is 2-bromo-3-methoxypyridine (prepared
according to methods known in the art from commercially available
2-bromo-3-hydroxy pyridine) or by the displacement of a 3-fluoro pyridine
intermediate 24 (in scheme V) with methoxide in methanol.

[0335] One of skill in the art would recognize that a variety of compounds
of the present invention may be prepared according to the general method
of Schemes I, II, III, IV, and V, according to methods known in the art,
and the synthetic Examples set forth below.

[0336] The compounds of this invention are potent inhibitors of gyrase and
Topo IV as determined by enzymatic assay. These compounds have also been
shown to have antimicrobial activity in an antimicrobial susceptibility
assay. The activity of a compound utilized in this invention as an
inhibitor of gyrase or Topo IV may be assayed in vitro, in vivo or in a
cell line according to methods known in the art. The details of the
conditions used for both the enzymatic and the antimicrobial
susceptibility assays are set forth in the Examples below.

[0337] According to another embodiment, the invention provides a
composition comprising a compound of this invention or a pharmaceutically
acceptable salt thereof and a pharmaceutically acceptable carrier,
adjuvant, or vehicle. The amount of compound in the compositions of this
invention is such that is effective to detectably inhibit gyrase, Topo
IV, or to measurably decrease bacterial quantity, in a biological sample
or in a patient. Preferably the composition of this invention is
formulated for administration to a patient in need of such composition.
Most preferably, the composition of this invention is formulated for oral
administration to a patient.

[0338] The term "biological sample", as used herein, includes, without
limitation, cell cultures or extracts thereof; biopsied material obtained
from a mammal or extracts thereof; and blood, saliva, urine, feces,
semen, tears, or other body fluids or extracts thereof.

[0339] Inhibition of gyrase and/or Topo IV activity in a biological sample
is useful for a variety of purposes that are known to one of skill in the
art. Examples of such purposes include, but are not limited to, blood
transfusion, organ-transplantation, biological specimen storage, and
biological assays.

[0340] The term "patient", as used herein, means an animal, preferably a
mammal, and most preferably a human.

[0342] The term "detectably inhibit", as used herein means a measurable
change in gyrase, or Topo IV, activity between a sample comprising said
composition and gyrase, or Topo IV, and an equivalent sample comprising
gyrase, or Topo IV in the absence of said composition.

[0343] As used herein, the term "measurably decrease bacterial quantity",
as used herein means a measurable change in the number of bacteria
between a sample containing said composition and a sample containing only
bacteria.

[0344] A "pharmaceutically acceptable salt" means any non-toxic salt of a
compound of this invention that, upon administration to a recipient, is
capable of providing, either directly or indirectly, a compound of this
invention or an inhibitorily active metabolite or residue thereof. As
used herein, the term "inhibitorily active metabolite or residue thereof"
means that a metabolite or residue thereof is also an inhibitor of gyrase
and/or Topo IV.

[0346] Salts derived from appropriate bases include alkali metal (e.g.,
sodium and potassium), alkaline earth metal (e.g., magnesium), ammonium
and N.sup.+ (C1-4 alkyl)4 salts. This invention also envisions
the quaternization of any basic nitrogen-containing groups of the
compounds disclosed herein. Water or oil-soluble or dispersible products
may be obtained by such quaternization.

[0347] The compositions of the present invention may be administered
orally, parenterally, by inhalation spray, topically, rectally, nasally,
buccally, vaginally or via an implanted reservoir. The term "parenteral"
as used herein includes subcutaneous, intravenous, intramuscular,
intra-articular, intra-synovial, intrasternal, intrathecal, intrahepatic,
intralesional and intracranial injection or infusion techniques.
Preferably, the compositions are administered orally, intraperitoneally
or intravenously. Sterile injectable forms of the compositions of this
invention may be aqueous or oleaginous suspension. These suspensions may
be formulated according to techniques known in the art using suitable
dispersing or wetting agents and suspending agents. The sterile
injectable preparation may also be a sterile injectable solution or
suspension in a non-toxic parenterally acceptable diluent or solvent, for
example as a solution in 1,3-butanediol. Among the acceptable vehicles
and solvents that may be employed are water, Ringer's solution and
isotonic sodium chloride solution. In addition, sterile, fixed oils are
conventionally employed as a solvent or suspending medium.

[0348] For this purpose, any bland fixed oil may be employed including
synthetic mono- or di-glycerides. Fatty acids, such as oleic acid and its
glyceride derivatives are useful in the preparation of injectables, as
are natural pharmaceutically-acceptable oils, such as olive oil or castor
oil, especially in their polyoxyethylated versions. These oil solutions
or suspensions may also contain a long-chain alcohol diluent or
dispersant, such as carboxymethyl cellulose or similar dispersing agents
that are commonly used in the formulation of pharmaceutically acceptable
dosage forms including emulsions and suspensions. Other commonly used
surfactants, such as Tweens, Spans and other emulsifying agents or
bioavailability enhancers which are commonly used in the manufacture of
pharmaceutically acceptable solid, liquid, or other dosage forms may also
be used for the purposes of formulation.

[0349] The pharmaceutically acceptable compositions of this invention may
be orally administered in any orally acceptable dosage form including,
but not limited to, capsules, tablets, aqueous suspensions or solutions.
In the case of tablets for oral use, carriers commonly used include
lactose and corn starch. Lubricating agents, such as magnesium stearate,
are also typically added. For oral administration in a capsule form,
useful diluents include lactose and dried cornstarch. When aqueous
suspensions are required for oral use, the active ingredient is combined
with emulsifying and suspending agents. If desired, certain sweetening,
flavoring or coloring agents may also be added.

[0350] Alternatively, the pharmaceutically acceptable compositions of this
invention may be administered in the form of suppositories for rectal
administration. These can be prepared by mixing the agent with a suitable
non-irritating excipient that is solid at room temperature but liquid at
rectal temperature and therefore will melt in the rectum to release the
drug. Such materials include cocoa butter, beeswax and polyethylene
glycols.

[0351] The pharmaceutically acceptable compositions of this invention may
also be administered topically, especially when the target of treatment
includes areas or organs readily accessible by topical application,
including diseases of the eye, the skin, or the lower intestinal tract.
Suitable topical formulations are readily prepared for each of these
areas or organs.

[0352] Topical application for the lower intestinal tract can be effected
in a rectal suppository formulation (see above) or in a suitable enema
formulation. Topically-transdermal patches may also be used.

[0353] For topical applications, the pharmaceutically acceptable
compositions may be formulated in a suitable ointment containing the
active component suspended or dissolved in one or more carriers. Carriers
for topical administration of the compounds of this invention include,
but are not limited to, mineral oil, liquid petrolatum, white petrolatum,
propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying
wax and water. Alternatively, the pharmaceutically acceptable
compositions can be formulated in a suitable lotion or cream containing
the active components suspended or dissolved in one or more
pharmaceutically acceptable carriers. Suitable carriers include, but are
not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl
esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

[0354] For ophthalmic use, the pharmaceutically acceptable compositions
may be formulated as micronized suspensions in isotonic, pH adjusted
sterile saline, or, preferably, as solutions in isotonic, pH adjusted
sterile saline, either with or without a preservative such as
benzylalkonium chloride. Alternatively, for ophthalmic uses, the
pharmaceutically acceptable compositions may be formulated in an ointment
such as petrolatum.

[0355] The pharmaceutically acceptable compositions of this invention may
also be administered by nasal aerosol or inhalation. Such compositions
are prepared according to techniques well-known in the art of
pharmaceutical formulation and may be prepared as solutions in saline,
employing benzyl alcohol or other suitable preservatives, absorption
promoters to enhance bioavailability, fluorocarbons, and/or other
conventional solubilizing or dispersing agents.

[0356] Most preferably, the pharmaceutically acceptable compositions of
this invention are formulated for oral administration.

[0359] Typically, the pharmaceutical compositions of this invention will
be administered from about 1 to 5 times per day or alternatively, as a
continuous infusion. Or, alternatively, the compositions of the present
invention may be administered in a pulsatile formulation. Such
administration can be used as a chronic or acute therapy. The amount of
active ingredient that may be combined with the carrier materials to
produce a single dosage form will vary depending upon the host treated
and the particular mode of administration. A typical preparation will
contain from about 5% to about 95% active compound (w/w). Preferably,
such preparations contain from about 20% to about 80% active compound.

[0360] When the compositions of this invention comprise a combination of a
compound of formula I or formula VII and one or more additional
therapeutic or prophylactic agents, both the compound and the additional
agent should be present at dosage levels of between about 10% to 80% of
the dosage normally administered in a monotherapy regime.

[0361] Upon improvement of a patient's condition, a maintenance dose of a
compound, composition or combination of this invention may be
administered, if necessary. Subsequently, the dosage or frequency of
administration, or both, may be reduced, as a function of the symptoms,
to a level at which the improved condition is retained when the symptoms
have been alleviated to the desired level, treatment should cease.
Patients may, however, require intermittent treatment on a long-term
basis upon any recurrence or disease symptoms.

[0362] As the skilled artisan will appreciate, lower or higher doses than
those recited above may be required. Specific dosage and treatment
regimens for any particular patient will depend upon a variety of
factors, including the activity of the specific compound employed, the
age, body weight, general health status, sex, diet, time of
administration, rate of excretion, drug combination, the severity and
course of the disease, and the patient's disposition to the disease and
the judgment of the treating physician.

[0363] Depending upon the particular condition, or disease, to be treated
or prevented, additional therapeutic agents, which are normally
administered to treat or prevent that condition, may also be present in
the compositions of this invention. As used herein, additional
therapeutic agents that are normally administered to treat or prevent a
particular disease, or condition, are known as "appropriate for the
disease, or condition, being treated". Such agents include, but are not
limited to, an antibiotic, an anti-inflammatory agent, a matrix
metalloprotease inhibitor, a lipoxygenase inhibitor, a cytokine
antagonist, an immunosuppressant, an anti-cancer agent, an anti-viral
agent, a cytokine, a growth factor, an immunomodulator, a prostaglandin,
an anti-vascular hyperproliferation compound, or an agent which increases
the susceptibility of bacterial organisms to antibiotics.

[0364] Agents that increase the susceptibility of bacterial organisms to
antibiotics are known. For example, U.S. Pat. No. 5,523,288, U.S. Pat.
No. 5,783,561 and U.S. Pat. No. 6,140,306 describe methods of using
bactericidal/permeability-increasing protein (BPI) for increasing
antibiotic susceptibility of gram-positive and gram-negative bacteria.
Agents that increase the permeability of the outer membrane of bacterial
organisms have been described by Vaara, M. in Microbiological Reviews
(1992) pp. 395-411, and the sensitization of gram-negative bacteria has
been described by Tsubery, H., et al, in J. Med. Chem. (2000) pp.
3085-3092.

[0365] According to another embodiment, the invention provides a method
for treating or lessening the severity of a bacterial infection in a
patient comprising the step of administering to said patient a
composition according to the present invention.

[0366] According to another embodiment, the invention provides a method of
inhibiting gyrase in a biological sample.

[0367] According to another embodiment, the invention provides a method of
inhibiting Topo IV in a biological sample.

[0368] According to another embodiment, the invention provides a method of
decreasing bacterial quantity in a biological sample.

[0369] According to another embodiment, the invention provides a method of
decreasing bacterial quantity in a biological sample, but further
comprising the step of contacting said biological sample with an agent
that increases the susceptibility of bacterial organisms to antibiotics.

[0372] In another embodiment, the pharmaceutical compositions and methods
of this invention will be useful generally for controlling bacterial
infections in vivo caused by the following the following organisms:
Streptococcus pneumoniae, Streptococcus pyogenes, Enterococcus faecalis,
Enterococcus faecium, Staphylococcus aureus, Coag. Neg. Staph, Bacillus
anthracis, Staphylococcus epidermidis, Staphylococcus saprophyticus, or
Mycobacterium tuberculosis.

[0373] The compositions and methods will therefore be useful for
controlling, treating or reducing the advancement, severity or effects of
nosocomial or non-nosocomial infections. Examples of nosocomial uses
include, but are not limited to, urinary tract infections, respiratory
infections such as pneumonia, surgical wound infections, and blood stream
infections (also known as bacteremia). Examples of non-nosocomial uses
include but are not limited to urinary tract infections, pneumonia,
prostatitis, skin and soft tissue infections, intra-abdominal infections,
and therapy for febrile neutropenic patients.

[0374] The term "pharmaceutically effective amount" refers to an amount
effective in treating or ameliorating a bacterial infection in a patient.
The term "prophylactically effective amount" refers to an amount
effective in preventing or substantially lessening a bacterial infection
in a patient.

[0375] The compounds of this invention may be employed in a conventional
manner for controlling bacterial infections levels in vivo and for
treating diseases or reducing the advancement or severity of effects that
are mediated by bacteria. Such methods of treatment, their dosage levels
and requirements may be selected by those of ordinary skill in the art
from available methods and techniques.

[0376] For example, a compound of this invention may be combined with a
pharmaceutically acceptable adjuvant for administration to a patient
suffering from a bacterial infection or disease in a pharmaceutically
acceptable manner and in an amount effective to lessen the severity of
that infection or disease.

[0377] Alternatively, the compounds of this invention may be used in
compositions and methods for treating or protecting individuals against
bacterial infections or diseases over extended periods of time. The
compounds may be employed in such compositions either alone or together
with other compounds of this invention in a manner consistent with the
conventional utilization of enzyme inhibitors in pharmaceutical
compositions. For example, a compound of this invention may be combined
with pharmaceutically acceptable adjuvants conventionally employed in
vaccines and administered in prophylactically effective amounts to
protect individuals over an extended period of time against bacterial
infections or diseases.

[0378] The compounds of formula I or formula VII may also be
co-administered with other antibiotics to increase the effect of therapy
or prophylaxis against various bacterial infections. When the compounds
of this invention are administered in combination therapies with other
agents, they may be administered sequentially or concurrently to the
patient. Alternatively, pharmaceutical or prophylactic compositions
according to this invention comprise a combination of a compound of
formula I or formula VII and another therapeutic or prophylactic agent.

[0379] The additional therapeutic agents described above may be
administered separately, as part of a multiple dosage regimen, from the
inhibitor-containing composition. Alternatively, these agents may be part
of a single dosage form, mixed together with the inhibitor in a single
composition.

[0380] In order that this invention be more fully understood, the
following examples are set forth. These examples are for the purpose of
illustration only and are not to be construed as limiting the scope of
the invention in any way.

EXAMPLES

Example 1

5-Bromo-1,3-difluoro-2-nitro-benzene

[0381] To a suspension of sodium perborate tetrahydrate (1.04 g, 5 mmol)
in acetic acid (20 mL), stirred at 55° C., was added a solution of
4-bromo-2,6-difluoroaniline in acetic acid (10 mL) over 1 hour in a
dropwise fashion. After stirring at 55° C. for an additional 3
hours, the solution was allowed to cool to room temperature and filtered.
The filtrate was poured into ice, and extracted twice with ethyl acetate.
The combined organic extracts were washed successively with
5×100-mL portions of water, brine, dried (MgSO4), and
concentrated in vacuo. The resulting residue was purified by column
chromatography over silica gel eluted with ethyl acetate:hexanes (1:20)
to afford 780 mg of the title compound as a tan solid. 1H NMR
(CDCl3) δ 7.32 (dt, 2H).

[0389] A mixture of 2-bromo-6-nitro-4-pyridin-3-yl-phenylamine (100 mg, 1
eq), 2-pyridylznic bromide (6 eq) and
tetrakis(triphenylphosphine)palladium (0.1 eq) in THF (10 mL) was heated
at 100° C. for 18 hours. The reaction was quenched with water (2
mL). The product was extracted with EtOAc (20×3). The combined
organic layer was then concentrated in vacuo and the residue was purified
by chromatography (Silica Gel, EtOAC) to afford the title compound (75
mg) as a yellow solid. (M+1) 293.

Example 10

3-Pyridin-2-yl-5-pyridin-3-yl-benzene-1,2-diamine

[0390] To a solution of 2-nitro-6-pyridin-2-yl-4-pyridin-3-yl-phenylamine
(75 mg, 0.26 mmol) in ethyl acetate (20 mL) was added 10% palladium on
carbon (50 mg). The resulting suspension was placed in a Parr
hydrogenation apparatus under 40 psi hydrogen gas while shaking at
ambient temperature for one hour. The catalyst was removed by filtration
and the filtrate concentrated in vacuo to afford compound the title
compound (50 mg, 0.19 mmol).

[0393] A 5 L flask was charged with the above depicted boronic acid as a
tetrahydrate (281.4 grams, 960 mmoles), 2-chloropyrimidine (100 g, 874
mmoles), NaHCO3 (146.8 grams, 1.746 moles), and Pd(PPh3)4
(10.0 grams, 8.72 mmoles). Water (1 L) and dimethoxyethane (1 L) were
added, and the mixture was heated slowly to 83° C. (internal
temperature) over a 1 hour period with overhead stirring. After ˜2
hours all solids had dissolved. The reaction was allowed to stir for 8
hours. The mixture was cooled to room temperature and stirred overnight
after which time a thick precipitate had formed. The crude mixture was
diluted with water (2 L) and stirred for an additional 2 hours after
which time the mixture was filtered and the solids were washed
sequentially with water, 0.1 N NaOH, and water again. The solids were
then dried under high vacuum at 50° C. to afford the title
compound (˜233 grams) as a tan powder.

Example 13

##STR00566##

[0394] N-(4-Bromo-2-pyrimidin-2-yl-phenyl)-2,2-dimethyl-propionamide

[0395] To a room temperature suspension of
2,2-dimethyl-N-(2-pyrimidin-2-yl-phenyl)-propionamide (˜117 grams,
437 mmoles) in acetic acid (1 L) was added bromine (67 mL, 1.31 moles) as
a solution in 100 mL of acetic acid over a 1 hour period. The
heterogenous mixture was stirred at room temperature for 5 hours over
which time a thick precipitate formed. The mixture was then poured over
ice, diluted with 1N Na2S2O3 (2 L), and stirred for 1
hour. The solids were filtered, resuspended in water (2 L), stirred for 1
hour, then filtered and washed with water again. The resulting solids
were pumped to dryness at 50° C., resuspended in HOAc (1 L), and
treated with bromine (22 mL, 430 mmoles) in acetic acid solution (20 mL)
over a 20 minute period. The resulting heterogenous mixture was stirred
for 5 hours, then quenched and treated as described above. The resulting
solids were vacuum dried at 50° C. to afford the title compound
(165 grams) as a tan powder.

Example 14

##STR00567##

[0396] N-(4-Bromo-2-nitro-6-pyrimidin-2-yl-phenyl)-2,2-dimethyl-propionami-
de

[0397] To a 5° C. suspension of
N-(4-bromo-2-pyrimidin-2-yl-phenyl)-2,2-dimethyl-propionamide (32.6
grams, 97.5 mmoles) in TFA (400 mL) was added 90% nitric acid (70 mL,
1.46 mmoles) over a 30 minute period. The mixture was then allowed to
warm to room temperature and stir for a total of 2 hours. The crude
reaction (now homogenous) was poured into ice producing a pasty mass. The
mixture was diluted to 2 L total volume with water, treated with 500 mL
of methanol, and vigorously stirred for 12 hours. The resulting solids
were filtered, washed with copious amounts of water, then vacuum dried at
50° C. to afford the title compound (29.9 grams, 81% yield) as a
tan powder.

Example 15

##STR00568##

[0398] 4-Bromo-2-nitro-6-pyrimidin-2-yl-phenylamine

[0399] A suspension of
N-(4-bromo-2-nitro-6-pyrimidin-2-yl-phenyl)-2,2-dimethyl-propionamide
(29.9 grams, 78.8 mmoles) in conc. HCl (200 mL) was refluxed for 8 hours.
The partially homogeneous crude reaction was then cooled to room
temperature, diluted with water (500 mL), and the resulting precipitate
was stirred for 1 hour. The solids were then filtered, washed with water,
and vacuum dried at 50° C. to afford the title compound (21.1
grams, 91% yield) as an orange powder.

[0401] A mixture of 4-bromo-2-nitro-6-pyrimidin-2-yl-phenylamine (1.82 g,
6.2 mmol), bis(pinacolato)diboron (3.144 g, 12.4 mmol),
PdCl2dppf2 (453 mg, 0.6 mmol) and KOAc (3.03 g, 31 mmol) in
dioxane (60 ml) was heated at 105° C. for 2.5 hours. The reaction
was filtered and washed with dichloromethane. The combined filtrates were
concentrated under vacuum and water (100 ml) was added to the residue.
Extraction with dichloromethane (3×50 ml), drying and concentration
gave a residue, which was washed with ether-hexane to afford the title
compound (2.07 g, 98%).

Example 17

##STR00570##

[0402] N-[2-(3-Fluoro-pyridin-2-yl)-phenyl]-2,2-dimethyl-propionamide

[0403] A 3 L flask was charged with the above depicted boronic acid as a
tetrahydrate (92.1 grams, 314 mmoles), chlorofluoropyridine (37.6 g, 286
mmoles), NaHCO3 (48.0 grams, 572 mmoles), and Pd(PPh3)4
(3.3 grams, 2.86 mmoles). Water (300 mL) and dimethoxyethane (300 mL)
were added, and the mixture was heated slowly to 83° C. (internal
temperature) over a 1 hour period with overhead stirring. After ˜2
hours all solids had dissolved. The reaction was allowed to stir for 10
hours. The mixture was cooled to room temperature and stirred overnight
after which time a thick gum had formed. The crude mixture was diluted
with water (2 L) and stirred for an additional 2 hours. The mixture was
then allowed to rest without stirring until the gum had settled to the
bottom of the flask. The liquid phase was removed via vacuum, then
replaced with 0.1 N NaOH and stirred for 15 minutes. The gum was allowed
to settle and the liquid removed via vacuum. The gum was then similarly
washed three times with water, then transferred to a 1 neck flask as an
acetone solution. The mixture was concentrated in vacuo and azeotroped
five times with ethyl acetate.

[0405] To a room temperature suspension of
N-[2-(3-fluoro-pyridin-2-yl)-phenyl]-2,2-dimethyl-propionamide (˜77
mmoles) in acetic acid (300 mL) was added bromine (12 mL, 228 mmoles) as
a solution in 50 mL of acetic acid over a 1 hour period. The heterogenous
mixture was stirred at room temperature for 5 hours over which time a
thick precipitate formed. The mixture was then poured over ice, diluted
with 1N Na2S2O3 (500 mL), and stirred for 1 hour. The
solids were filtered, re-suspended in water (2 L), stirred for 1 hour,
then filtered and washed with water again. The resulting solids were
pumped to dryness at 50° C., re-suspended in HOAc (400 mL), and
treated with bromine (4 mL, 76 mmoles) in acetic acid solution (20 mL)
over a 20 minute period. The resulting heterogenous mixture was stirred
for 5 hours, then quenched and treated as described above. The resulting
solids were vacuum dried at 50° C. to afford the title compound
(19.1 grams, 72%) as a tan powder.

[0407] To a suspension of
N-[4-bromo-2-(3-fluoro-pyridin-2-yl)-phenyl]-2,2-dimethyl-propionamide
(6.45 grams, 18.4 mmoles) in TFA (100 mL) and TFAA (25.5 mL, 183.6
mmole), at 0° C., was added a TFA solution (30 mL) of 90% fuming
nitric acid (2.46 mL, 55.1 mmoles) over a 45 minute period. The mixture
was then stirred at 0° C. for a total of 4 hours. The crude
reaction (now homogenous) was poured into ice producing a pasty mass. The
mixture was diluted to 500 mL total volume with water, treated with 50 mL
of methanol, and vigorously stirred for 12 hours. The resulting solids
were filtered, washed with copious amounts of water, then dried in vacuo
at 50° C. to afford the title compound (6.1 grams, 82% yield) as a
tan powder.

[0413] To a solution of 2-(3,5-difluoro-2-nitro-phenyl)-pyrimidine (1.5 g,
6.32 mmoles) in dioxane (10 mL) was added tBuNH2 (6.6 mL, 63.24
mmoles) at room temperature. The mixture was heated to 100° C. in
a sealed tube for 10 hours. The mixture was then cooled to room
temperature, poured into water, and the solids stirred for 1 hour. The
mixture was filtered, solids washed with water until filtrate was clear.
The crude product was then diluted in MeOH, 6N HCl added, and the
resulting mixture heated at reflux for 3 hours. The reaction was cooled
to room temperature and poured into ice. The resulting heterogeneous
mixture was warmed to room temperature, filtered, solids washed with
water until filtrate ran clear, and dried in vacuo to afford the title
compound (1.33 g, 90%) as an orange powder. 1H NMR (dmso-d6,
500 MHz): 8.87 (d, 2H); 7.52 (dd, 1H); 7.08 (dd, 1H); 6.86 (dd, 1H); 6.60
(s, 2H).

[0422] We have prepared other compounds of formula I by methods
substantially similar to those described in Schemes I through IV,
Examples 1 through 26, and by methods known in the art. The
characterization data for these compounds is summarized in Table 3 below
and includes LC/MS (observed) and 1H NMR data.

[0423]1H NMR data is summarized in Table 3 below wherein 1H NMR
data was obtained at 500 MHz in deuterated DMSO, unless otherwise
indicated, and was found to be consistent with structure. Compound
numbers correspond to the compound numbers listed in Table 2.

[0424]1H NMR data is also summarized in Table 3a below wherein
1H NMR data was obtained at 500 MHz in the deuterated solvents
indicated therein, and was found to be consistent with the structure.
Compound numbers correspond to the compound numbers listed in Table 2a.

[0425] The ATP hydrolysis activity of DNA gyrase was measured by coupling
the production of ADP through pyruvate kinase/lactate dehydrogenase to
the oxidation of NADH. This method has been described previously (Tamura
and Gellert, 1990, J. Biol. Chem., 265, 21342).

[0426] ATPase assays are carried out at 30° C. in buffered
solutions containing 100 mM TRIS pH 7.6, 1.5 mM MgCl2, 150 mM KCl.
The coupling system contains (final concentrations) 2.5 mM phosphoenol
pyruvate, 200 μM nicotinamide adenine dinucleotide (NADH), 1 mM DTT,
30 ug/ml pyruvate kinase, and 10 ug/ml lactate dehydrogenase. 40
nanomolar enzyme (374 kDa Gyr A2B2 subunit from Staphylococcus aureus)
and a DMSO solution of the inhibitor to a final concentration of 4% are
added and the reaction mixture is allowed to incubate for 10 minutes at
30° C. The reaction is then started by the addition of ATP to a
final concentration of 0.9 mM and the rate of NADH disappearance at 340
nanometers is measured over the course of 10 minutes. The Ki values
are determined from rate versus concentration profiles.

[0427] Compounds of the present invention were found to inhibit gyrase. In
certain embodiments, compounds of the present invention inhibit gyrase
with a Ki value of less than 50 nM in the above assay.

Example 28

Topo IV ATPase Assay

[0428] The conversion of ATP to ADP by Topo4 enzyme is coupled to the
conversion of NADH to NAD+ and measured by the change in absorbance at
340 nm. Topo4 is incubated with inhibitor (4% DMSO final) in buffer for
10 minutes at 30° C. Reaction is initiated with ATP and rates are
monitored continuously for 20 minutes at 30° C. on a Molecular
Devices SpectraMAX plate reader. The inhibition constant, Ki, is
determined from plots of rate vs. [Inhibitor] fit to the Morrison
Equation for tight binding inhibitors.

[0431] Compounds of the present invention were found to inhibit TopoIV. In
certain embodiments, compounds of the present invention inhibit TopoIV
with a Ki value of less than 50 nM in the above assay.

Example 29

Susceptibility Testing in Liquid Media

[0432] Compounds of this invention were also tested for antimicrobial
activity by susceptibility testing in liquid media. Such assays were
performed within the guidelines of the latest NCCLS document governing
such practices: "M7-A5 Methods for dilution Antimicrobial Susceptibility
Tests for Bacteria that Grow Aerobically; Approved Standard--Fifth
Edition (2000)". Other publications such as "Antibiotics in Laboratory
Medicine" (Edited by V. Lorian, Publishers Williams and Wilkins, 1996)
provide essential practical techniques in laboratory antibiotic testing.
Essentially, several discrete bacterial colonies of Staphylococcus
aureus, Streptococcus pneumoniae, Enterococcus faecalis, Enterococcus
faecium, E. coli, Haemophilus influenzae, Staphylococcus epidermidis, or
Staphylococcus saprophyticus (3 to 7) from a freshly streaked plate were
transferred to an appropriate rich broth medium such as MHB (Mueller
Hinton broth), supplemented where appropriate for the more fastidious
organisms. This was grown overnight to high density followed by a 1 or
2-thousand-fold dilution to give an inoculation density of between
5×105 and 5×106 CFU per mL. Alternatively, the
freshly picked colonies can be incubated at 37° C. for about 4 to
8 hours until the culture equals or exceeds a turbidity of a 0.5
McFarland standard (approximately 1.5×108 cells per mL) and
diluted to give the same CFU per mL as above. In a more convenient
method, the inoculum was prepared using a commercially available
mechanical device (the BBL PROMPT System) that involves touching five
colonies directly with a wand, containing crosshatch grooves at its
bottom, followed by suspension of the bacteria in an appropriate volume
of saline. Dilution to the appropriate inoculum cell density was made
from this cell suspension. The broth used for testing consists of MHB
supplemented with 50 mg per L of Ca2+ and 25 mg per L of Mg2+.
In the case of Staphylococcus aureus, the relative serum binding was
measured in assays containing 50% frozen human serum in cation-adjusted
Mueller Hinton broth. Standard dilution panels of control antibiotics
were made and stored as in the NCCLS standard M7-A5, the dilution range
typically being in the 128 μg per mL to 0.015 μg per mL (by 2-fold
serial dilution). The test compounds were dissolved and diluted fresh for
experimentation on the same day; the same or similar ranges of
concentration as above being used. The test compounds and controls were
dispensed into a multiwell plate and test bacteria added such that the
final inoculation was approximately 5×104 CFU per well and the
final volume was 100 μL. The plates were incubated at 35° C.
overnight (16 to 20 hours) and checked by eye for turbidity or
quantitated with a multiwell plate reader. The endpoint minimal
inhibitory concentration (MIC) is the lowest concentration of drug at
which the microorganism tested (Staphylococcus aureus, Streptococcus
pneumoniae, Enterococcus faecalis, Enterococcus faecium, E. coli,
Haemophilus influenzae, Staphylococcus epidermidis, or Staphylococcus
saprophyticus) does not grow under the test conditions. Such
determinations were also compared to the appropriate tables contained in
the above two publications to ensure that the range of antibacterial
activity is within the acceptable range for this standardized assay.

[0434] Compounds of this invention were also tested for antimicrobial
activity against certain resistant bacterial colonies by susceptibility
testing in liquid media. Such assays were performed within the guidelines
of the latest NCCLS document governing such practices: "M7-A5 Methods for
dilution Antimicrobial Susceptibility Tests for Bacteria that Grow
Aerobically; Approved Standard--Fifth Edition (2000)". Other publications
such as "Antibiotics in Laboratory Medicine" (Edited by V. Lorian,
Publishers Williams and Wilkins, 1996) provide essential practical
techniques in laboratory antibiotic testing. Essentially, several
discrete bacterial colonies of Methicillin resistant Staphylococcus
aureus, Fluoroquinolone resistant Staphylococcus aureus, Vancomycin
intermediate resistant Staphylococcus aureus, Linezolid resistant
Staphylococcus aureus, Penicillin resistant Streptococcus pneumoniae,
Macrolide resistant Streptococcus pneumoniae, Fluoroquinolone resistant
Streptococcus pneumoniae, Vancomycin resistant Enterococcus faecalis,
Linezolid resistant Enterococcus faecalis, Fluoroquinolone resistant
Enterococcus faecalis, Vancomycin resistant Enterococcus faecium,
Linezolid resistant Enterococcus faecium, Fluoroquinolone resistant
Enterococcus faecium, Ampicillin resistant Enterococcus faecium,
Macrolide resistant Haemophilus influenzae, Beta-lactam resistant
Haemophilus influenzae, Fluoroquinolone resistant Haemophilus influenzae,
Beta-lactam resistant Moraxella catarrhalis, Methicillin resistant
Staphylococcus epidermidis, Methicillin resistant Staphylococcus
epidermidis, Vancomycin resistant Staphylococcus epidermidis,
Fluoroquinolone resistant Staphylococcus epidermidis, Macrolide resistant
Mycoplama pneumoniae, Isoniazid resistant Mycobacterium tuberculosis, or
Rifampin resistant Mycobacterium tuberculosis (3 to 7) from a freshly
streaked plate were transferred to an appropriate rich broth medium such
as MHB (Mueller Hinton broth), supplemented where appropriate for the
more fastidious organisms. This was grown overnight to high density
followed by a 1 or 2-thousand-fold dilution to give an inoculation
density of between 5×105 and 5×106 CFU per mL.
Alternatively, the freshly picked colonies can be incubated at 37°
C. for about 4 to 8 hours until the culture equals or exceeds a turbidity
of a 0.5 McFarland standard (approximately 1.5×108 cells per
mL) and diluted to give the same CFU per mL as above. In a more
convenient method, the inoculum was prepared using a commercially
available mechanical device (the BBL PROMPT System) that involves
touching five colonies directly with a wand, containing crosshatch
grooves at its bottom, followed by suspension of the bacteria in an
appropriate volume of saline. Dilution to the appropriate inoculum cell
density was made from this cell suspension. The broth used for testing
consists of MHB supplemented with 50 mg per L of Ca2+ and 25 mg per
L of Mg2+. Standard dilution panels of control antibiotics were made
and stored as in the NCCLS standard M7-A5, the dilution range typically
being in the 128 μg per mL to 0.015 μg per mL (by 2-fold serial
dilution). The test compounds were dissolved and diluted fresh for
experimentation on the same day; the same or similar ranges of
concentration as above being used. The test compounds and controls were
dispensed into a multiwell plate and test bacteria added such that the
final inoculation was approximately 5×104 CFU per well and the
final volume was 100 μL. The plates were incubated at 35° C.
overnight (16 to 20 hours) and checked by eye for turbidity or
quantitated with a multiwell plate reader. The endpoint minimal
inhibitory concentration (MIC) is the lowest concentration of drug at
which the microorganism tested does not grow under the test conditions.

[0436] While we have described a number of embodiments of the present
invention, it is apparent that our basic constructions may be altered to
provide other embodiments that utilize the products and processes of this
invention.